Genome sequencing data have been accumulating exponentially. The detection and analysis of a tremendous amount of genetic information require new rapid, highly-efficient techniques of hybridization and sequencing. The development of high-through genechip technology has dramatically enhanced our ability in male infertility research. Current applications of genechip technology in male infertility include the study of testis genes, the analysis of spermatozoon mRNA, the study on cell genital toxicity, the diagnosis and treatment of male infertility. This review summarizes the present situation in male infertility research and the potential clinical application.
Animals ; Homeodomain Proteins ; genetics ; Humans ; Infertility, Male ; diagnosis ; genetics ; therapy ; Male ; Oligonucleotide Array Sequence Analysis ; RNA, Messenger ; analysis

OBJECTIVETo analyze HOXD13 gene in polydactyly in dispersion type of Fujian Han population in order to know whether there is mutation in HOXD13.

METHODSAll members were evaluated physically and radlologically. Genomic DNA was extracted from peripheral blood of the patients who were treated from Dec. 2012 to Apr. 2013, their parents, grandparents, and normal volunteers from our department. The polymerase chain reaction ( PCR) , agarose gel electrophoresis and DNA sequence analysis were adopted to analyze HOXD13 from six cases with polydactyly and forty normal volunteers.

RESULTSAll patients had no family history. A heterozygous synonymous mutation, c. 291 C > T( p. A60A), was detected in exon 1 of the HOXD13 Gene in five of the polydactyly patients. Similar mutation was not detected in one brachy dactyly patient and the forty normal volunteers.

CONCLUSIONA heterozygous synonymous mutation, c.291C > T (p. A60A), of the HOXD13 gene may be related with polydactyly in dispersion type of Chinese han population.

China ; Exons ; Heterozygote ; Homeodomain Proteins ; genetics ; Humans ; Mutation ; Polydactyly ; genetics ; Polymerase Chain Reaction ; Sequence Analysis, DNA ; Transcription Factors ; genetics

OBJECTIVETo investigate the effects of lentivirus-mediated RNA interference targeting HOXA10 gene on the proliferation, apoptosis and morphology of leukemic cell line U937.

METHODSFour different shRNA plasmids were designed and built to interfere with HOXA10 gene. The four interference plasmids were transfected into 293T cells with the HOXA10 over expression plasmid and then the RNAi efficiency of the four interference plasmids was determined by Western blot. The best one was chosen to transfect 293T cells with lentiviral helping plasmids to produce packaged lentivirus (lenti-shHOXA10). U937 cells were divided into interference group (lenti-shHOXA10), negative control group and untreated group. After infection with the packaged lentivirus, infection efficiency of lentivirus for U937 was detected by flow cytometry, and the expression of HOXA10 gene mRNA and protein was detected by real-time PCR and Western blot. Cell survival was determined by MTT assay. Apoptosis rate was detected by flow cytometry.

RESULTSLentiviral-shRNA vector of HOXA10 gene was successfully constructed. Compared with the negative control and untreated groups, mRNA level of HOXA10 decreased by (92.3±1.3)%, protein levels decreased by 91.1%, and the inhibition rate of U937 cells [(43.9±0.7)%] increased in the interference group (P<0.05). Wright's staining showed that the ratio of karyon to cytoplasm was reduced and mitotic phase was rare in the interference group. Apoptosis rate in the interference group [(27.1±1.4)%] was significantly higher than in the negative [(19.4±1.9)%] and untreated groups [(5.5±1.3)%] (P<0.05).

CONCLUSIONSLentivirus mediated RNAi can reduce the expression level of HOXA10, effectively inhibit proliferation and promote apoptosis of U937 cells. HOXA10 gene is expected to become a new target for the treatment of leukemia at gene level.

Apoptosis ; Cell Proliferation ; Gene Silencing ; Homeodomain Proteins ; antagonists & inhibitors ; genetics ; Humans ; Lentivirus ; genetics ; RNA Interference ; Sequence Analysis, DNA ; U937 Cells

Animals ; Glucose ; pharmacology ; Homeodomain Proteins ; analysis ; genetics ; physiology ; Immunohistochemistry ; Insulin ; secretion ; Male ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Trans-Activators ; analysis ; genetics ; physiology

Hypermethylation of the homeobox (HOX) gene promoter leads to decreased expression of the gene during tumor development and is thought to be correlated with the clinical outcome in leukemia. In this study, we performed pyrosequencing to quantify the methylation level of HOXA5 genes in the bone marrow samples obtained from 50 patients with AML and 19 normal controls. The methylation percentage of HOXA5 in AML patients (median=65.4%, interquartile range=35.9-72.3%) was higher than that of HOXA5 in control patients (median=43.1%, interquartile range=36.7-49.6%, Mann-Whitney U test, P=0.012). The patients of the AML group who had a high methylation percentage (>70%) had a good prognosis with a 3-yr overall survival (OS) of 82.5%, whereas the patients with a low methylation percentage (< or =70%) showed a 3-yr OS of 40.5% (P=0.048). Cox proportional hazards regression showed that the methylation percentages of HOXA5 were independently associated with the 3-yr OS of AML patients, regardless of their karyotypes. We propose that the quantification of HOXA5 methylation by pyrosequencing may be useful for predicting short-term prognosis in AML. However, the limitations of our study are the small sample size and its preliminary nature. Thus, a larger study should be performed to clearly determine the relationships among HOXA5 methylation levels, cytogenetics, and prognosis in AML patients.
Adult ; Aged ; *DNA Methylation ; Female ; Homeodomain Proteins/*genetics ; Humans ; Leukemia, Myeloid, Acute/genetics/*mortality ; Male ; Middle Aged ; Prognosis ; Promoter Regions, Genetic ; ROC Curve ; Sequence Analysis, DNA ; Survival Rate

BACKGROUND/AIMS: The caudal-related homeobox transcription factor, Cdx2, plays an important role in proliferation and differentiation of intestinal epithelial cells. Its expression is confined to normal and neoplastic intestinal epithelium. We evaluated Cdx2 expression in advanced colorectal cancers to determine the correlation between Cdx2 expression and clinicopathologic characteristics. METHODS: Four hundreds twenty consecutive colorectal cancers were included in the study. Cdx2 expression was investigated by immunohistochemistry using tissue microarrays constructed from surgically resected specimens. 145 invasive breast cancers, normal tissues from gastric mucosa, liver, lung, kidney and ovary were used as control. Nuclear staining was considered to be positive and the result was divided into 3 categories. RESULTS: In the colorectal cancers, Cdx2 was expressed in 380 of 420 (90.5%) cases, and 349 of 380 (83%) cases showed strong and diffuse staining and 31 of 420 (7.5%) cases showed weakly positive staining. Forty patients (9.5%) of colorectal cancer were negative for Cdx2. All of the invasive breast cancers and all non-neoplastic control tissues except the regions of intestinal metaplasia in gastric mucosa, which showed strong Cdx2 expression, were negative for Cdx2. Loss of Cdx2 expression was observed more frequently in cases with deeper invasion (p<0.05), lymph node metastasis (p<0.05), poor histologic differentiation (p<0.001), and distant metastasis (p<0.05). CONCLUSIONS: Cdx2 could be a highly sensitive marker to detect metastasis from intestine and might be useful as a novel prognostic marker in colorectal cancers.
Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms/*metabolism/pathology ; Female ; Homeodomain Proteins/*metabolism ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Tumor Markers, Biological/analysis

OBJECTIVETo study the clinical features and to identify homeobox D13 (HOXD13) gene mutation of the affected individuals in a Chinese synpolydactyly (SPD) kindred.

METHODSClinical data and peripheral blood samples of SPD family members were obtained through field investigation. For every member of this pedigreeìthe fragment containing mutational hot spots of HOXD13 was amplified by PCR for mutation screening. To examine whether there is any other mutation within coding sequence of HOXD13, exon 1 and exon 2 of HOXD13 were also amplified by PCR. All the amplified fragments were electrophoresed on 2% agarose gels and then the mutant fragments were electrophoresed on 5% polyacrylamide gels to be separated. Purified PCR products of normal and selected mutant alleles were directly sequenced.

RESULTSComparing the HOXD13 coding sequence of the affected individuals with HOXD13 sequence in the GenBank and with that of the unaffected, an inserted segment coding 8 alanine residues within HOXD13 was found segregating with the disorder. This mutation is also termed polyalanine expansion. The 8-alanine expansion can be interpreted as a reduplication of normal alanines 5-12.

CONCLUSIONThe results suggest that synpolydactyly in this kindred may be caused by polyalanine expansion in HOXD13.

Base Sequence ; China ; DNA Mutational Analysis ; Female ; Homeodomain Proteins ; genetics ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Pedigree ; Polymerase Chain Reaction ; Syndactyly ; genetics ; Transcription Factors ; genetics

OBJECTIVETo construct a high effective eukaryotic expressing plasmid PcDNA 3.1-MSX-2 encoding Sprague-Dawley rat MSX-2 gene for the further study of MSX-2 gene function.

METHODSThe full length SD rat MSX-2 gene was amplified by PCR, and the full length DNA was inserted in the PMD1 8-T vector. It was isolated by restriction enzyme digest with BamHI and Xhol, then ligated into the cloning site of the PcDNA3.1 expression plasmid. The positive recombinant was identified by PCR analysis, restriction endonudease analysis and sequence analysis. Expression of RNA and protein was detected by RT-PCR and Western blot analysis in PcDNA3.1-MSX-2 transfected HEK293 cells.

RESULTSSequence analysis and restriction endonudease analysis of PcDNA3.1-MSX-2 demonstrated that the position and size of MSX-2 cDNA insertion were consistent with the design. RT-PCR and Western blot analysis showed specific expression of mRNA and protein of MSX-2 in the transfected HEK293 cells.

CONCLUSIONSThe high effective eukaryotic expression plasmid PcDNA3.1-MSX-2 encoding Sprague-Dawley Rat MSX-2 gene which is related to craniofacial development can be successfully reconstructed. It may serve as the basis for the further study of MSX-2 gene function.

Animals ; Cloning, Molecular ; Gene Expression ; Genes, Homeobox ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Plasmids ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Sequence Analysis, DNA

Diagnostic Techniques, Respiratory System ; Female ; Follow-Up Studies ; Homeodomain Proteins ; analysis ; therapeutic use ; Humans ; Hypoventilation ; physiopathology ; Male ; Sleep Apnea Syndromes ; diagnosis ; drug therapy ; therapy

OBJECTIVETo study the changes of Hes-1, the target gene of Notch signaling pathway, and its relationship with airway inflammation and remodeling in a rat model of asthma.

METHODSForty-eight rats were randomly divided into an asthma group and a control group. The rats in the asthma group were sensitized and challenged by ovalbumin (OVA), and normal saline was used in the control group. Two groups were further divided into 3 subgroups according to time points after challenging, i.e. 4 weeks, 8 weeks and 12 weeks (n=8 rats each). Pathological changes of lungs were observed by light microscopy and the thickness of bronchial smooth muscle layer (Wam) was measured. The levels of IL-4 and INF-γ in rat serum and bronchoalveolar lavage fluids (BALF) were measured using ELISA. Expression levels of Hes-1 protein and mRNA were determined by immunohistochemistry and quantitative real-time PCR respectively.

RESULTSTogether with the extension of challenging, the Wam of rats in the asthma group increased, a decrease of INF-γ level and an increase of IL-4 level in serum and BALF were also observed, and the differences were statistically significant compared with those in the corresponding control group (P<0.05). Hes-1 protein and mRNA levels also increased gradually after OVA challenging and were higher than those in the control group (P<0.05). The levels of Hes-1 protein and mRNA were positively correlated with Wam and IL-4 in serum and BALF, but were inversely correlated with INF-γ in serum and BALF (P<0.05).

CONCLUSIONSLevels of Hes-1 protein and mRNA increased, which were closely related with the levels of airway inflammatory factors and remodeling of airway smooth muscle. Hes-1 may play an important role in the pathogenesis of asthma.

Airway Remodeling ; Animals ; Asthma ; etiology ; Basic Helix-Loop-Helix Transcription Factors ; analysis ; genetics ; physiology ; Disease Models, Animal ; Homeodomain Proteins ; analysis ; genetics ; physiology ; Interferon-gamma ; analysis ; Interleukin-4 ; analysis ; Male ; Rats ; Rats, Sprague-Dawley ; Transcription Factor HES-1